Measuring chip for surface plasmon resonance biosensor and method for producing the same

Abstract

An objective of the present invention is to provide a measuring chip for a surface plasmon resonance sensor that can detect a small amount of target substances in high sensitivity. The present invention provides a measuring chip for a surface plasmon resonance sensor comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a biologically active substance immobilized on the surface of said plasma polymerization layer.

1 . A measuring chip for a surface plasmon resonance sensor comprising 
 a metal layer and    one or more plasma polymerization layers formed on said metal layer.    
   
   
       2 . The measuring chip of  claim 1 , wherein said metal layer comprises gold, platinum or silver.  
   
   
       3 . The measuring chip of  claim 1  further comprising a nucleic acid immobilized on the surface of at least one plasma polymerization layers.  
   
   
       4 . The measuring chip for a surface plasmon resonance sensor of  claim 1  further comprising 
 a non-immune protein immobilized on the surface of at least one plasma polymerization layer.    
   
   
       5 . The measuring chip for a surface plasmon resonance sensor of  claim 1  further comprising a non-immune protein which is avidin, streptoavidin, biotin or a receptor.  
   
   
       6 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , further comprising 
 an immunoglobulin-binding protein immobilized on the surface of at least one plasma polymerization layer.    
   
   
       7 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , further comprising an immunoglobulin-binding protein which is protein A, protein G, or a rheumatoid factor.  
   
   
       8 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , further comprising 
 a sugar-binding protein immobilized on the surface of at least one plasma polymerization layer.    
   
   
       9 . The measuring chip for a surface plasmon resonance sensor of  claim 1  further comprising a sugar-binding protein which is a lectin.  
   
   
       10 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a sugar-recognizing sugar chain immobilized on the surface of said plasma polymerization layer.  
   
   
       11 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , comprising a metal layer, one or more plasma polymerization layers formed on said metal layer, and a fatty acid or a fatty acid ester immobilized on the surface of said plasma polymerization layer.  
   
   
       12 . The measuring chip for a surface plasmon resonance sensor of  claim 1  further comprising a fatty acid or fatty acid ester which is stearic acid, alachidic acid, behenic acid, ethyl stearate, ethyl arachidate, or ethyl behanate.  
   
   
       13 . The measuring chip for a surface plasmon resonance sensor of  claim 1  further comprising a polypeptide or oligopeptide having ligand-binding activity immobilized on the surface of said at least one plasma polymerization layer.  
   
   
       14 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , further comprising a polypeptide or oligopeptide which is produced by genetic engineering or chemical synthesis.  
   
   
       15 . The measuring chip of  claim 1  further comprising an optically transparent substrate on which said metal layer is formed.  
   
   
       16 . A measuring chip according to  claim 1 , wherein said one or more plasma polymerization layer(s) comprises a compound having one or more substitutents selected from the group consisting of —COOH, —CHO, —SH, —NH 2 , —OH, ═NH, —CONH 2 , —NCO, —CH═CH 2 , ═C═O, and  
     
       
         
         
             
             
         
       
     
   
   
       17 . A measuring chip of  claim 1 , comprising two or more plasma polymerization layers.  
   
   
       18 . A measuring chip of  claim 1 , wherein at least one plasma polymerization layer is formed from a monomer material containing nitrogen.  
   
   
       19 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , wherein said plasma polymerization layer is formed from a compound containing nitrogen which is CH 3 —(CH 2 ) n —NH 2  (wherein n is an integer from 1 to 6) and/or NH 2 —(CH 2 ) n —NH 2  (wherein n is an integer from 1 to 6).  
   
   
       20 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , comprising a plasma polymerization layer which is produced from a compound containing nitrogen selected from the group consisting of pyridine, ethylenediamine, hexamethylenediamine, n-propylamine, monoethylamine, triethylamine, diethylamine, allylamine, acrylamide, aniline, acrylonitrile, 1,2,4-triazole, 5-amino-1H-tetrazole, propargylamine and acetonitrile.  
   
   
       21 . The measuring chip of  claim 1 , wherein at least one plasma polymerization layer is formed from a monomer material containing sulfur.  
   
   
       22 . The measuring chip according to  claim 1 , wherein at least one plasma polymerization layer is formed from a compound containing sulfur selected from the group consisting of dimethyl sulfide, methyl disulfide, ethanethiol, ethanedithiol, thiophen, mercaptoethanol and dithreitol.  
   
   
       23 . The measuring chip according to  claim 1 , wherein said plasma polymerization layer is formed from a monomer compound containing a halogen.  
   
   
       24 . The measuring chip according to  claim 1 , wherein said at least one plasma polymerization layer is formed from a monomer material having one or more groups selected from the group consisting of —COON, —CHO, —SH, —NH 2 , —OH, ═NH, —CONH 2 , —NCO, —CH═CH 2 , ═C═O and  
     
       
         
         
             
             
         
       
     
   
   
       25 . The measuring chip according to  claim 1 , wherein said monomer material of a plasma polymerization layer is a compound having —C═CCH 2 OH.  
   
   
       26 . The measuring chip according to  claim 1 , wherein said plasma polymerization layer is formed from a monomer material which is a carbohydrate compound comprising C and H.  
   
   
       27 . The measuring chip according to  claim 1 , wherein said plasma polymerization layer is formed from a monomer material which is an organic metal compound.  
   
   
       28 . A measuring chip according to  claim 27 , wherein said organic metal compound is an organic silicon compound.  
   
   
       29 . A measuring chip according to  claim 28 , wherein said organic silicon compound is selected from the group consisting of tetramethylsilane, tetramethyldisiloxane, hexamethyldisiloxane, hexamethyldisilazane, hexamethylcyclotrisilazane, dimethylaminotrimethylsilane, trimethylvinylsilane, tetramethoxysilane, aminopropyltriethoxysilane, octadecyldiethoxymethylsilane, hexamethyldisilane, and divinyltetramethyldisiloxane.  
   
   
       30 . the measuring chip according to  claim 1 , wherein plasma treatment is applied to form said plasma polymerization layer with a polymeric or non-polymeric monomer.  
   
   
       31 . A measuring chip according to  claim 30  wherein said non-polymeric monomer material is selected from the group consisting of nitrogen, ammonium, hydrazine, hydrogensulfide, hydrogendisulfide, oxygen, hydrogen, water, halogen gas, and rare gas.  
   
   
       32 . The measuring chip according to  claim 1 , wherein said plasma polymerization layer is formed from a mixture of two or more different types of monomers.  
   
   
       33 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , further comprising an immune protein or enzyme immobilized on the surface of said plasma polymerization layer, wherein said plasma polymerization layer comprises a monomer material selected from the group consisting of pyridine, triethylamine, diethylamine, allylamine, acrylamide, aniline, acrylonitrile, 1,2,4-triazole, 5-amino-1H-tetrazole, and acetonitrile.  
   
   
       34 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , further comprising an immune protein or enzyme immobilized on the surface of said plasma polymerization layer, wherein said plasma polymerization layer comprises a monomer material selected from the group consisting of a compound containing sulfur, an oxygen containing compound, a carbon containing compound, a compound containing a halogen, an organic metal compound, an organic silicon compound, and a carbohydrate compound containing C and H.  
   
   
       35 . A measuring chip according to  claim 33 , wherein said immune protein is an antibody.  
   
   
       36 . measuring chip according to  claim 33 , wherein said immune protein is a Fab fragment of an antibody.  
   
   
       37 . A measuring chip according to  claim 33 , wherein said immune protein is an F(ab)2 fragment of an antibody.  
   
   
       38 . A measuring chip according to  claim 3 , wherein said nucleic acid is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
   
   
       39 . A measuring chip according to  claim 4 , wherein said non-immune protein is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
   
   
       40 . A measuring chip according to  claim 6 , wherein said immunoglobulin-binding protein is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
   
   
       41 . A measuring chip according to  claim 8 , wherein said sugar-binding protein is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
   
   
       42 . A measuring chip according to  claim 10 , wherein said sugar-recognizing sugar chain is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
   
   
       43 . A measuring chip according to  claim 11 , wherein said fatty acid or fatty acid ester is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
   
   
       44 . A measuring chip according to  claim 13 , wherein said polypeptide or oligopeptide is immobilized on said plasma polymerization layer through a cross-linking reagent or a condensation reagent.  
   
   
       45 . The measuring chip according to  claim 38 , wherein said cross-linking reagent is one or more compounds selected from the group consisting of glutaraldehyde, periodic acid, N-succinimidyl-2-maleimidoacetic acid, N-succinimidyl-4-maleimidobutyric acid, N-succinimidyl-6-maleimidohexanic acid, N-succinimidyl-4-maleimidomethylcyclohexan-1-carboxylic acid, N-sulfosuccinimidyl-4-maleimidomethylcyclohexane-1-carboxylic acid, N-succinimidyl-4-maleimidomethylbanzoic acid, N-succinimidyl-3-maleimidobenzoic acid, N-sulfosuccinimidyl-3 maleimidobenzoic acid, N-succinimidyl-4-maleimidophenyl-4-butyric acid, N-sulfosuccinimidyl-4-maleimidophenyl-4-butyric acid, N,N′-oxydimethylene-dimaleimide, N,N′-o-phenylene-dimaleimide, N,N′-m-phenylene-dimaleimide, N,N′-p-phenylene-dimaleimide, N,N′-hexamethylene-dimaleimide, N-succinimidylmaleimidocarboxylic acid, N-succinimidyl-S-acetylmercaptoacetic acid, N-succinimidyl-3-(2-pyridyldithio)propionate, S-acetylmercaptosuccinic anhydride, methyl-3-(4′-dithiopyridyl)propionimidate, methyl-4-mercaptobutylimidate, methyl-3-mercaptopropionimidate, iminothiolene, o-carboxymethyl-hydroxylamine, azodiphenylpilmaleido, bis(sulfosuccinimidyl)sperate, 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene, 4,4′-difluoro-3,3′-dinitrodiphenylsulfon, 1,5-difluoro-2,4-dinitrobenzene, p-phenylenediisothiocyanate, dimethyladipimidate, dimethylpimelimidate, dimethylsuberimidate, p-azidophenacylbromide, p-azidophenylglyoxal, N-hydroxysuccinimidyl-4-azidobenzoate, 4-fluoro-3-nitrophenylazide, methyl-4-azidobenzoimidate, N-5-azido-2-nitrobenzoyloxysuccinimide, N-succinimidyl-6-(4′-azido-2′-nitrophenylamino)hexanoate, 1,4-benzoquinone, N-succinimidyl-3-(2′-pyridyldithio)propionate, N-(4-maleimidobutyloxy)sulfosuccinimide sodium salt, N-(6-maleimidocaproyloxy)sulfosuccinimide sodium salt, N-(8-maleimidocaproyloxy)sulfosuccinimide sodium salt, N-( 11-maleimidoundecanoyloxy)sulfosuccinimide sodium salt, N-[2-(1-piperazinyl)ethyl] maleimide bichloric acid, bisdiazobenzidine, hexamethylenediisocyanate, toluenediisocyanate, hexamethylenediisothiocyanate, N,N′-ethylenebismaleinimide, N,N′-polymethylenebisiodoacetamide, 2,4-dinitrobenzenesulfonate sodium salt, and diazo compounds; or said condensation reagent is one or more compounds selected from the group consisting of carbodiimide derivatives represented by RN═C═NR (or R′), N-hydroxysuccinimide, tri-n-butylamine, butyl chloroformate, and isobutyl isocyanide.  
   
   
       46 . A measuring chip according to  claim 1 , wherein said immune protein or enzyme is immobilized on said plasma polymerization layer through a cross-linking reagent or a water-soluble condensation reagent.  
   
   
       47 . A measuring chip according to  claim 46 , wherein said cross-linking reagent is one or more compounds selected from the group consisting of glutaraldehyde, N-succinimidyl-4-maleimidomethylbanzoic acid, N-succinimidyl-3-maleimidobenzoic acid, N-succinimidyl-4-maleimidophenyl-4-butyric acid, N,N′-oxydimethylene-dimaleimide, N,N′-m-phenylene-dimaleimide, N,N′-p-phenylene-dimaleimide, N,N′-hexamethylene-dimaleimide, N-succinimidylmaleimidocarboxylic acid, N-succinimidyl-S-acetylmercaptoacetic acid, N-succinimidyl-3-(2-pyridyldithio)propionate, S-acetylmercaptosuccinic anhydride, methyl-3-(4′-dithiopyridyl)propionimidate, methyl-4-mercaptobutylimidate, methyl-3-mercaptopropionimidate, iminothiolene, o-carboxymethyl-hydroxylamine, azodiphenylpilmaleido, bis(sulfosuccinimidyl)sperate, 4,4′-diisothiocyano-2,2′-disulfonic acid stilbene, 4,4′-difluoro-3,3′-dinitrodiphenylsulfon, 1,5-difluoro-2,4-dinitrobenzene, p-phenylenediisothiocyanate, dimethyladipimidate, dimethylpimelimidate, dimethylsuberimidate, p-azidophenacylbromide, p-azidophenylglyoxal, N-hydroxysuccinimidyl-4-azidobenzoate, 4-fluoro-3-nitrophenylazide, methyl-4-azidobenzoimidate, N-5-azido-2-nitrobenzoyloxysuccinimide, N-succinimidyl-6-(4′-azido-2′-nitrophenylamino)hexanoate, 1,4-benzoquinone, N-succinimidyl-3-(2′-pyridyldithio)propionate, bisdiazobenzidine, hexamethylenediisocyanate, toluenediisocyanate, hexamethylenediisothiocyanate, N,N′-ethylenebismaleinimido, N,N′-polymethylenebisiodoacetoamide, and diazo compounds; or said condensation reagent is one or more compounds selected from the group consisting of carbodiimide derivatives represented by RN═C═NR (or R′), N-hydroxysuccinimide, tri-n-butylamine, butyl chloroformate, and isobutyl isocyanide.  
   
   
       48 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , further comprising a substance immobilized on the surface of said plasma polymerization layer, and an additional plasma polymerization layer or plasma-treated layer formed on said plasma polymerization layer.  
   
   
       49 . A measuring chip according to  claim 48 , wherein said substance to be immobilized is selected from the group consisting of a non-immune protein, an immunoglobulin-binding protein, a sugar-binding protein, a sugar-recognizing sugar chain, a fatty acid or a fatty acid ester, a polypeptide or oligopeptide having ligand-binding activity, an immune protein, and an enzyme.  
   
   
       50 . The measuring chip for a surface plasmon resonance sensor of  claim 1 , further comprising a substance immobilized on said plasma polymerization layer through a hydrophobic bond.  
   
   
       51 . The measuring chip according to  claim 50 , wherein said substance to be immobilized is selected from the group consisting of a a non-immune protein, an immunoglobulin-binding protein, a sugar-binding protein, a sugar-recognizing sugar chain, a fatty acid or a fatty acid ester, a polypeptide or oligopeptide having ligand-binding activity, an immune protein, and an enzyme.  
   
   
       52 . A method for producing a measuring chip for a surface plasmon resonance sensor comprising: 
 forming a metal layer on an optically transparent substrate,    forming one or more plasma polymerization layers on said metal layer, and immobilizing a physiologically active substance on the surface of said plasma polymerization layer.    
   
   
       53 . The method of  claim 52 , comprising immobilizing a non-immune protein on the surface of said plasma polymerization layer.  
   
   
       54 . The method of  claim 52  comprising immobilizing an immunoglobulin-binding protein on the surface of said plasma polymerization layer.  
   
   
       55 . The method of  claim 52  comprising A immobilizing a sugar-binding protein on the surface of said plasma polymerization layer.  
   
   
       56 . The method of  claim 52  comprising a sugar-recognizing sugar chain on the surface of said plasma polymerization layer.  
   
   
       57 . The method of  claim 52  comprising _ immobilizing a fatty acid or fatty acid ester on the surface of said plasma polymerization layer.  
   
   
       58 . The method of  claim 52  comprising immobilizing a polypeptide or oligopeptide having a ligand binding capability on the surface of said plasma polymerization layer.  
   
   
       59 . A method according to  claim 52  wherein the plasma polymerization layer is formed by a plasma-treatment using a monomer material.  
   
   
       60 . The method of  claim 52  comprising 
 immobilizing an immune protein or enzyme on the surface of said plasma polymerization layer.    
   
   
       61 . A measuring cell for a surface plasmon resonance sensor comprising a measuring chip according to  claim 1 .  
   
   
       62 . A measuring cell according to  claim 61 , wherein said chip is optically analyzed.  
   
   
       63 . A surface plasmon resonance biosensor comprising a measuring chip according to  claim 1 .  
   
   
       64 . A surface plasmon resonance biosensor comprising a measuring cell according to  claim 61.